Biological Activity of the Alternative Promoters of the Dictyostelium discoideum Adenylyl Cyclase A Gene
Fig 4
Expression of aggregation-specific genes in complemented acaA-mutant strains.
Cells from the wild-type AX4 strain (WT), the acaA-mutant strain (acaA-KO) or the mutant strain transformed with acaA expression vectors under control of the acaA promoters 1 (KO/Pr1::acaA), 2 (KO/Pr2::acaA) or 3 (KO/Pr3::acaA) were deposited on Nitrocellulose filters soaked on the phosphate-based PDF Buffer to induce development. RNAs were isolated from cells collected either before placing them on the filters (time 0) or at different times of development (3, 6, 9, 12, 15, 18, 21 and 24 hours). The expression of the genes coding for cAMP receptor 1 (carA, upper panel) and the cell-adhesion molecule TgrC5 (tgrC5, lower panel) was determined by reverse transcription and quantitative PCR. Expression of the mitochondrial large ribosomal RNA was used as internal control. Expression at each developmental stage was normalized to that of non-starved cells (time 0) for each gene and strain. Average values and standard deviations of three experiments made in triplicate are represented.